ABSTRACT
The use of quinolone for treatment of rickettsial diseases remains controversial. Recent clinical studies suggest that quinolone is not as effective as others in patients with rickettsial diseases including scrub typhus, although the mechanism is not well understood. In this study, we evaluated the mutation in gyrA associated with quinolone resistance. We prospectively enrolled scrub typhus patients, collected blood samples and clinical data from October, 2010 to November, 2011. Among the 21 patients enrolled, one initially received ciprofloxacin for 3 days but was switched to doxycycline due to clinical deterioration. We obtained the gyrA gene of Orientia tsutsugamushi from 21 samples (20 Boryong strain, 1 Kato strain) and sequenced the quinolone resistance-determining region. All of 21 samples had the Ser83Leu mutation in the gyrA gene, which is known to be associated with quinolone resistance. This suggests that quinolones may be avoided for the treatment of serious scrub typhus.
Subject(s)
Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Amino Acid Sequence , Anti-Bacterial Agents/therapeutic use , Bacterial Proteins/genetics , Ciprofloxacin/therapeutic use , DNA Gyrase/genetics , Doxycycline/therapeutic use , Drug Resistance, Bacterial , Genotype , Molecular Sequence Data , Mutation , Orientia tsutsugamushi/classification , Phylogeny , Prospective Studies , Scrub Typhus/drug therapy , Sequence Alignment , Sequence Analysis, DNAABSTRACT
There are several antigenic variants of Orientia tsutsugamushi. The 56-kDa type-specific antigen (TSA) is responsible for the antigenic variation. Nucleotide sequences of the 56-kDa TSA obtained from 44 eschar samples of Korean scrub typhus patients and from 40 representative strains retrieved from the GenBank database were analyzed phylogenetically. Clinical patient data were assessed based on the genotyping results. Of the 44 nucleotide sequences, 32 (72.7%) clustered with the Boryong genotype, which is the major genotype in Korea. Eleven nucleotide sequences (25%) clustered with the Kawasaki genotype, not identified in Korea until 2010. One nucleotide sequence was consistent with the Karp genotype. The clinical course of the patients infected with each genotype showed no differences. Diagnostic performance of the immunofluorescence assay (IFA) using the 56-kDa TSA from Gilliam, Karp and Boryong as test antigens were not different for the Boryong and Kawasaki genotypes. Although Boryong is still the predominant genotype, the results suggest that Kawasaki genotype is quite prevalent in Chungbuk province of Korea.
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Bacterial Proteins/genetics , Base Sequence , DNA, Bacterial/analysis , Databases, Genetic , Genotype , Open Reading Frames , Orientia tsutsugamushi/classification , Phylogeny , Republic of Korea , Scrub Typhus/diagnosis , Sequence Analysis, DNAABSTRACT
There are several antigenic variants of Orientia tsutsugamushi. The 56-kDa type-specific antigen (TSA) is responsible for the antigenic variation. Nucleotide sequences of the 56-kDa TSA obtained from 44 eschar samples of Korean scrub typhus patients and from 40 representative strains retrieved from the GenBank database were analyzed phylogenetically. Clinical patient data were assessed based on the genotyping results. Of the 44 nucleotide sequences, 32 (72.7%) clustered with the Boryong genotype, which is the major genotype in Korea. Eleven nucleotide sequences (25%) clustered with the Kawasaki genotype, not identified in Korea until 2010. One nucleotide sequence was consistent with the Karp genotype. The clinical course of the patients infected with each genotype showed no differences. Diagnostic performance of the immunofluorescence assay (IFA) using the 56-kDa TSA from Gilliam, Karp and Boryong as test antigens were not different for the Boryong and Kawasaki genotypes. Although Boryong is still the predominant genotype, the results suggest that Kawasaki genotype is quite prevalent in Chungbuk province of Korea.
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Bacterial Proteins/genetics , Base Sequence , DNA, Bacterial/analysis , Databases, Genetic , Genotype , Open Reading Frames , Orientia tsutsugamushi/classification , Phylogeny , Republic of Korea , Scrub Typhus/diagnosis , Sequence Analysis, DNAABSTRACT
The seroprevalence of various Orientia tsutsugamushi (OT) strains among Malaysian patients with suspected scrub typhus infections was determined using an indirect immunoperoxidase (IIP) assay. IgG against a single OT strain were detected in six sera (3 Karp, 1 Gilliam and 2 TC586), whereas IgM antibodies against a single OT strain (Gilliam) were noted in 3 sera (Gilliam). IgG reactive to all OT strains were present in 33 (47.1%) of the 70 sera and IgM reactive to all OT strains were present in 22 (78.6%) of the 28 sera. The fact that most sera were reactive to multiple OT strains suggests that group-specific antigens are involved in scrub typhus infections, whereas very few were due to strain-specific epitopes present on these strains. Peak IgG and IgM titers were noted more frequently against Gilliam, Karp, and TA763 strains: this suggests that these strains may be the commonest infecting strains among Malaysian patients. Two predominant OT polypeptides consistently reacted with patients' sera were the 70 kDa and 56 kDa proteins.
Subject(s)
Antigens, Bacterial/analysis , Humans , Immunoblotting , Immunoenzyme Techniques/methods , Immunoglobulin G/blood , Immunoglobulin M/blood , Malaysia/epidemiology , Orientia tsutsugamushi/classification , Scrub Typhus/epidemiology , Seroepidemiologic StudiesABSTRACT
The genotype of Orientia tsutsugamushi DNA from mites in the Xisa archipelago of China were identified. A natural focus of tsutsugamushi disease in the archipelago was found. The DNA sequence that codes for the 56 kDa protein of O. tsutsugamushi was amplified by nested polymerase chain reaction (N-PCR). The purified positive products were cloned into a pGEM-T vector and sequenced. The DNA sequence was compared with various sequences on the internet for sequence homology. A 507 bp DNA fragment encoding the 56 kDa protein was amplified from the samples. The sequence homology was 85% (Karp strain), 68% (Gilliam strain), 65% (Kato strain), and 67% (Yonchon strain). Orientia tsutsugamushi is carried by the mites of the Xisa archipelago; the main genotype is the Karp strain.
Subject(s)
Animals , Bacterial Typing Techniques/methods , China , DNA, Bacterial/analysis , Genotype , Humans , Mites/microbiology , Orientia tsutsugamushi/classification , Polymorphism, Restriction Fragment Length , Reference ValuesABSTRACT
No abstract available.
Subject(s)
Humans , Animals , Antibodies, Bacterial/blood , Antigens, Bacterial/blood , Mites/microbiology , Orientia tsutsugamushi/classification , Scrub Typhus/diagnosisABSTRACT
R. tsutsugamushi strains found in chiggers collected from 3 different sites throughout Thailand were antigenically characterized. Monotypic infections were observed in 76.7% of the chiggers. Karp was the most predominant strain, followed by TA716, TA763, TA686 and Kato. This study represents the first confirmed report of R. tsutsugamushi infection in 8 different chigger species in Thailand.